IJCBS, 22(2022): 79-85

International Journal of Chemical and Biochemical Sciences

(ISSN 2226-9614)

Journal Home page: www.iscientific.org/Journal.html

© International Scientific Organization

Antibacterial and antifungal activities of polygon mint essential oils

Abdellah LAQHAILI*1, Reda CHAROF2, Zakaria MENNANE2, Sanaâ SAOAIBI3,
Malika SABIHA1, Ayoub NAJEM1, Asmae El-YAHYAOUI4, Souhayla Latifi2, Abdelkbir
BELLAOUCHOU1, Ahmed SAOAIBI3, El Mostapha LOTFI4, Mahjouba MOSADDAK1,
Abdelhak HAKIKI1, Abdellah GUENBOUR1, Younes ZAID1,5
1
Laboratory of Materials, Nanotechnology, and Environment (Natural substances and Flash Thermolysis),
Department of Chemistry. faculty of Sciences rabat, Mohammed V University, P.O. Box. 1014 Agdal-Rabat, Morocco
2
Laboratory of Epidemic Diseases Department of Medical Bacteriology National Institute of Hygiene Rabat Morocco
3
Laboratory of Chimie Appliquée des Matériaux, Department of Chemistry, Faculty of Sciences, Mohammed V
University in Rabat, Morocco.
4
Laboratory of Spectroscopy, Molecula Modeling, Materials, Nanomaterials, Water and Environment, Materials for
Environment Team, ENSAM, Mohammed V University in Rabat, Morocco.
5
Botany Laboratory, Department of Biology, Faculty of Sciences, Mohammed V University, Rabat, Morocco.

Abstract

In this study, we worked on a medicinal aromatic plant widely used in Morocco, namely, Mentha pulegium L. of the
Lamiaceae family (Labiatae). This plant was characterized by the Azilal region (Middle Atlas) and another from the Gharb region,
then was placed in a well-ventilated place under light and shade to dry it, followed by extraction by the method of hydrodistillation
for the extraction of essential oil from pennyroyal. The spectral analyzes by (CPG) and (CPG-MS) show that the oil extracted before
and after maceration is rich, especially in Pulegone for the two plants of the two regions. A study of the effect of maceration on
essential oil yield showed that the defined plant essential oil yield of the Azilal region is RHE= 2.32% without maceration and
RHE= 2.33% after maceration, on the other hand, the essential oil yield of the plant in the Gharb region is RHE= 2.27% before
maceration and RHE= 2.30% after maceration. The antibacterial activity of the essential oil of Mentha pulgium from the two regions
confirmed that the essential oil extracted from the plant indicated in the region of Azilal has a very significant zone of inhibition
compared to that of the region of Gharb.

Keywords: Mentha pulgium, essential oil, maceration, aromatic plant

Short communication *Corresponding Author, e-mail: [email protected]

1. Introduction
disease, inflammation, and allergy problems [4]. Nowadays
Medicinal plants play a significant role in drug discovery traditional medicine or treatment using natural plants is still
[1]. It is reported that at least 119 compounds derived from limited [5]. For this, research has been conducted on the study
90 plant species can be considered important drugs against of natural and biologically active products [6]. These
most diseases [2]. Unfortunately, until today researchers have products are chemical substances of organism origin not used
not found effective drugs [3] for several diseases namely, in the primary metabolism [7]. Indeed, the selection of the
cancers, viral infections such as HIV, hepatitis C, infections plant to be studied is a crucial factor in the ultimate success
by bacteria and fungi, diseases cardiovascular of the extraction of biologically active agents [8].
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In Morocco, mint is widely used for flavoring tea and in In order to study the antibacterial and antifungal activity
traditional medicine. For a long time, Morocco had been of the essential oil, tests by the agar diffusion method on 25
trying to develop the peppermint industry and the Gharb strains, namely Enterobacteriaceae (Enterococcus focalize.
plain, with its fertile alluvial soil, contributed to an essential Escherichia coli, Enterobacter Freundii; Serratia marcesens,
oil production estimated at 5 tons in 1946. Other regions are etc.), Non-Enterobacteriaceae (Pseudomonas aerogenosa,
favorable such as the south of tiflet and khemisset, with acenitobacterbaumanii); Gram-positive Cocci
Màaziz [9][10]. Mints are plants of great perennial (staphylococcal and streptococcal), Gram-positive Bacillus
herbaceous vigor with a characteristic odor that leaves an (Listeria spp) and for yeasts we tested Candida albicans.
impression of freshness, and they have been used for
therapeutic purposes since the 16th century [11]. In therapy, 3. Results and Discussions
mint is used against fever, weakness, cough, nausea, stomach
aches, melancholy, chest diseases, hysteria, and visual 3.1. Study of maceration on yield
disturbances [12][13], it also presents medicinal properties,
for example, antioxidant [14] antifungal [15], antibacterial The essential oil yield for the Azilal region (average atlas)
[16], anticancer activity and with radioactive potential [17]. is RHE= 2.32% without maceration is 2.33% after
Mint is also used against parasites [18]; the stems and flowers maceration, on the other hand for the Gharb region the
of mint are burned to drive away fleas from mattresses and essential oil yield is RHE= 2.27% without maceration is
domestic animals [19][20]. 2.30% after maceration.

In this context, the objective of our work is to study the 3.2. Analysis by GC and GC/MS
maceration and the antibacterial activity of the essential oil
extracted from the Mentha pulegium L. plant of the Qualitative and quantitative analyses of essential oils
Lamiaceae family (Labiaceae), harvested from two different were made using gas chromatography, and gas
regions, the Azilal region, and the Gharb region. The chromatography coupled with mass spectrophotometry for
identification of the compounds presents in the essential oil the two plants, respectively.
to be studied was carried out using GC/MS.
3.3. Chemical composition of the essential oil before
2. Materials and methods maceration

2.1. Materials Figure 1 gives us the percentage of the main majority

products with their respective retention time. Indeed, the
The studied plant Mentha pulgium L. was harvested in the retention times can give information on the nature of the
region of Azilal (middle atlas), and placed in a well-ventilated molecules and the areas of the peaks provide a relative
place sheltered from light and shade. After drying, the plant quantification.
was ready for extraction. From the chromatogram, it can be seen that the majority
of the product corresponds to the peak which appears for a
2.2. Methods retention time equal to 19.78 min. Analysis by GC and GC-
MS allows us to confirm that our oil is rich in Polygon.
• Extraction of essential oils:
3.4. Chemical composition of the essential oil after
The extraction was carried out using the hydrodistillation maceration
method. A quantity of 40% of the plant is completely This chromatogram gives us the main majority
immersed in 60% of distilled water, and to homogenize the compounds of the extracted gasoline. The results give us the
boiling of the mixture some grains of pumice stone has been main majority compounds with their respective retention
added. The heating is maintained at a gentle temperature for times. :2-Octyne, 1,1-diethoxy-. The two methods of analysis
8 hours. the vapors are condensed in the cooler and the enabled us to identify the main compounds of the essential oil
essential oil and water are separated by difference in density. extracted after maceration. It was noted that the majority of
products appear for the following peaks: 19.80; 17.4; we also
• Yield variation over time. note the formation of new products. This allows us to say that
Yield= (Mxp / Mth) × 100. the maceration of the plant influences the chemical
composition.
RHE= (mass of HE / mass of dry plant) x 100
3.5. Antibacterial and antifungal activity
2.3. Characterization
According to figure 3 and table 1, it can be seen that the
An analysis by gas chromatography coupled with mass essential oil of Middle Atlas mint (HE (1)) has a very
spectrometry (GPC/MS) was carried out to identify and significant activity because this oil inhibits almost the
calculate the percentages of the constituents present in the majority of bacteria (80%). This is remarkable for
essential oil extracted from the mint. The calculations were Enterobacteriaceae, Staphylocossus, and Listeria spp. On the
carried out according to the internal standard method. other hand, it remains ineffective on Streptococcus
pnomoniae and Panteo.

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Table 1: Results of antibacterial and antifungal activities

Bacterial strains Inhibition diameters (mm)

Type of germ
Essential Oil N°1 Essential Oil N°2
Dilution Dilution Dilution Dilution
Brute Brute
1/2 1/4 1/2 1/4

Enterococcus foecalis 16 11 12 14 12 14

Yersina ATCC 23 19 9 - - -

Serratia marcesens 16 12 9 - - -

Salmonella Entritidis 19 11 15 12 - -
Esherichiacoli beta
lactamas à spectre 20 19 21 14 11 -
Élargie (E.C BLSE)
Escherichia coli
24 18 16 12 - -
O157

Shigella spp 22 22 19 12 - 11

ENTEROBACTERIE
11 11 - - -
Enterobacteries
COLOACAE
PROTEUS spp 20 16 14 - -

Klebsiella spp 24 18 11 14 - -

Marganellaspp 28 26 20 13 8 11

SALMONELLE spp 18 15 10 - - -

Klebsiella BLSE 15 12 - - - -

Panteospp - - - - - -

Salmonella Groupe
19 20 15 11 - -
C

Staphylococcus
28 22 14 12 8 10
Aureus ATCC

Streptococcus
- - - - - -
COCCI pneumoniae
GRAM+
Staphylococcus
22 20 15 - - -
epedermis

Streptococcus groupe 16 11 - - - -
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Psudomonas
- - - - - -
aeruginosa ATCC
No Enterobacteriaceae
Acinetobactere
28 20 18 21 16 17
bounanii

BACILLE G+ Listeria spp 11 - - - - -

RT: 4.00 - 45.03 SM: 15G

RT: 19.78 NL:
100 4.50E7
95 TIC F: MS
F443GC02
90 RT: 17.03
85
80
75
70
65
60
55 RT: 16.61
50
45
RT: 14.29
40
35
30
25
20
15
10
RT: 22.98
5 RT: 12.12
RT: 6.93
RT: 26.64 31.21 36.44 RT: 41.58
0
5 10 15 20 25 30 35 40 45
Time (min)

Figure 1: Chromatogram of the relative abundance as a function of the time of the essential oil before maceration.

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 IJCBS, 22(2022): 79-85

RT: 0.00 - 45.03 SM: 7G

19.80 NL:
100 4.68E7
TIC F: MS
80 F443GC02
Relative Abundance

60 17.04

40

20
22.98
8.75 12.12 26.64 31.21 35.45 39.64
0
0 5 10 15 20 25 30 35 40 45
Time (min)

Figure 2: Chromatogram of the relative abundance as a function of time of the essential oil
extracted after maceration

a: Shaphylococcus aureus ATCC

b: Salmonelle entritidis

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 IJCBS, 22(2022): 79-85

c: Escherichia coli O157 d : PROTEUS

e: Klebsiella
f: Salmonella Groupe C

Figure 3: Action of essential oils extracted on different bacteria

The essential oil of pennyroyal from the Gharb region of Salmonella, Yersina ATCC) as well as the germs responsible
Morocco (HE (2)) significantly inhibited Enterobacteriaceae, for urinary tract infections and nosocomial infections (ESBL
especially the strains: Enterococcus, Escherichia coli, Klebsiella, Escherichia coli O157, etc.). The differences in
Klebsella spp. For Cocci Gram+; HE (2) inhibited only inhibition can be explained by the chemical variability of the
Shaphylococcus aureus and remains ineffective for two extracts, which depends on the nature of the soil, the
Streptococcus pneumoniae, Staphylococcus epedermis, degree of pollution, and the nature of the climate.
Streptococcus group D. For Non-Enterobacteriaceae, HE (2)
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